The invention relates to a method for reconstructing a three-dimensional fluorescence optical tomography image to examine a plate-shaped object having a first face and an opposite second face and comprising fluorophores, said method comprising:                a first illumination step for a sequential illumination of the first face of the object with a fluorophore excitation light from a plurality of lighting points, the first illumination step comprising a succession of illumination phases from a lighting point associated with each phase, and        a first acquisition step for a sequential acquisition of a first series of images, an image of said series being formed, at each illumination phase of the first face, by a detector comprising a plurality of detection points simultaneously detecting light emitted by the second face of the object.        